恶性疟血涂片见大滋养体1例

本文报告1例非洲输入性非典型恶性疟病例, 其血涂片显微镜下可见较多大滋养体期原虫, 部分虫体可见棕黄色 色素颗粒, 呈点状或团块状, 虫体形态与间日疟原虫、 卵形疟原虫相似, 经PCR基因检测确诊为恶性疟。非重症恶性疟病 例外周血中出现大滋养体期原虫较为罕见, 镜检时应注意与其他疟原虫鉴别。     

1例血涂片查到恶性疟原虫各期形态的报告

多年来,国内有关间日疟原虫多形性报道很多,包括多重感染,多核,条带状滋养体,成熟裂殖体较小,裂殖体数量少,配子体比例较高等非典型形态特征。但对于恶性疟原虫形态研究的报道较少,现将1例少见的血涂片恶性疟原虫形态观察报告如下。     

间日疟原虫红细胞外期体外培养的研究

用人肝癌细胞在体外培养间日疟原虫红细胞外期获得成功，在国内首次建立肝癌细胞—间日疟原虫红细胞外期体外培养系统。在无菌条件下解剖受感染的斯氏按蚊唾腺，制成子孢子悬液，并接种到肝癌细胞中进行体外培养，７ｄ后在培养物中可见红细胞外期裂殖体。第１０ｄ，在培养物中加入正常人Ｏ型红细胞，继续培养，１４ｄ后分离红细胞制成涂片，可见大量环状体和早期大滋养体。提示此种肝癌细胞可作为我国间日疟原虫红细胞外期体外培养的宿主细胞。     

服用四环素后疟原虫形态发生改变的1例间日疟病例

目的 提高疟原虫镜检人员对使用过具有抗疟作用的四环素药物后形态发生改变的疟原虫虫种鉴定能力,为疟疾病例的治疗和疫点处置提供科学依据,巩固疟疾消除成果。方法 对2022年9月2日大理大学第一附属医院接诊的1例间日疟患者胡某某进行流行病学调查,采集其血液样本进行吉姆萨染色,在显微镜油镜下观察并描述疟原虫形态、被疟原虫寄生的红细胞大小和颜色,鉴定虫种;分别用RDT和巢式PCR方法对血样进行复核。结果 胡某某于2022年8月25日起出现发冷、发热、出汗,每天不规则发热,全身酸痛,患者认为再次感染恙虫病,每日口服四环素500 mg,连服4 d;2022年8月29日胡某某入住大理州洱源县人民医院,2022年9月2日转大理大学第一附属医院。镜检疟原虫发现各期疟原虫均发生明显变形：小滋养体出现1环2核、边缘寄生现象,被其他期疟原虫寄生的红细胞显示颜色变淡、胀大、细胞内有细小而多的薛氏小点;大滋养体变成典型的带状,被寄生的红细胞明显胀大并且颜色变淡;配子体呈椭圆形;部份被疟原虫寄生的红细胞边缘呈毛刺状或突起,但部份未被疟原虫寄生的红细胞也有边缘呈毛刺状或突起。综合分析后鉴定为间日疟,经云南省寄生虫病防治...     

间日疟原虫环子孢子蛋白基因片段的扩增应用于诊断间日…

根据间日疟原虫环子孢子蛋白（ＣＳＰ）基因序列，设计并合成间日疟原虫特异性引物一对，采用聚合酶锭反应技术，进行间日疟原虫感染的检测。结果：（１）从间日疟原虫患者的全血ＤＮＡ中扩增出约７７２ｂｐ的基因片段，经限制性内切酶切，证实为间日疟原虫ＣＳＰ基因片段；（２）与间日疟原虫亲缘上近的三株恶性疟原虫，弓形虫，利什曼原虫和正常人全血核酸抽提物经扩增后均未见特异性的扩增条带；（３）该检测体系可检测出间日疟原     

PCR检测疟疾混合感染的现场应用研究

目的 评价PCR检测技术在间日疟与恶性疟混合感染区诊断疟疾的现场应用价值。方法 采集海南省疟疾混合感染流行区304份滤纸干血滴样本，根据红内期疟原虫SSUrRNA基因序列，设计合成3条引物．采用PCR技术在同一反应体系中扩增出间日疟原虫和恶性疟原虫不同的DNA片段，检测现场所采样本中的间日疟原虫或恶性疟原虫DNA；同时与镜检法进行比较。结果 在304份样本中，PCR法阳性15份，其中间日疟7份．恶性疟8份；镜检法阳性11份，其中间日疟6份，恶性疟5份。镜检阳性的样本PCR均为阳性；镜检阴性而PCR阳性的4份样本，其扩增产物经限制性酶切鉴定，证实为间日疟原虫或恶性疟原虫DNA。结论 此PCR检测体系灵敏、特异．对诊断或鉴别诊断间日疟原虫和恶性疟原虫混合感染具有实用价值。     

PCR检测疟疾混合感染的现场应用研究

目的　评价PCR检测技术在间日疟与恶性疟混合感染区诊断疟疾的现场应用价值。　方法　采集海南省疟疾混合感染流行区 3 0 4份滤纸干血滴样本 ,根据红内期疟原虫SSUrRNA基因序列 ,设计合成 3条引物 ,采用PCR技术在同一反应体系中扩增出间日疟原虫和恶性疟原虫不同的DNA片段 ,检测现场所采样本中的间日疟原虫或恶性疟原虫DNA ;同时与镜检法进行比较。　结果　在 3 0 4份样本中 ,PCR法阳性 15份 ,其中间日疟 7份 ,恶性疟 8份 ;镜检法阳性11份 ,其中间日疟 6份 ,恶性疟 5份。镜检阳性的样本PCR均为阳性 ;镜检阴性而PCR阳性的 4份样本 ,其扩增产物经限制性酶切鉴定 ,证实为间日疟原虫或恶性疟原虫DNA。　结论　此PCR检测体系灵敏、特异 ,对诊断或鉴别诊断间日疟原虫和恶性疟原虫混合感染具有实用价值。     

间日疟原虫与异种疟原虫之间的红内期交叉反应原分析

本文用１％ＮＰ－４０分别提取间日疟原虫、恶性疟原虫、食蟹猴疟原虫、伯氏疟原虫及约氏疟原虫中溶性抗原，用间日疟病血清及两株抗间疟红细胞内原虫单克隆抗体６Ｈ７和２Ｃ３对上述抗原进行免疫印迹分析。结果表明，间日疟原虫与异种疟原虫之间有一系列交叉反应原，其中食触猴疟原虫有１４条蛋白带可被间日疟病人血清识别，多于另外三种疟原虫。五的虫共有的７９ｋＤ蛋白均可被间日疟病人血清识别。此外，间日疟病人血清还识别正常     

湘北地区间日疟原虫形态特征的初步观察

对湘北地区60例间日疟病例疟原虫形态计量研究的结果表明,双核疟原虫和双重感染的阳性率分别为50.0％和48.3％。双核幼年滋养体和发育中滋养体分别占同期疟原虫的5.1％和1.3％,两者之比为3.9:1,与杨柏林等1982年的观察结果基本一致。说明双核疟原虫或双重感染在间日疟原虫中相当普遍。本文同时与云南思茅地区6例同种疟原虫作了比较。     

套式PCR检测蚊体内疟原虫的敏感性与特异性

目的：分析套式聚合酶链反应技术检测蚊体内疟原虫的敏感性与特异性。方法：采用２对针对间日疟原虫小亚单位核糖体核糖核酸基因（ＳＳＵｒＤＮＡ）的特异引物，利用套式ＰＣＲ技术，从蚊体ＤＮＡ样本中，扩增间日疟原虫ＳＳＵｒＤＮＡ片段，进行间日疟原虫的检测。结果：扩增产物经琼脂糖凝胶电泳分析，可见扩增出特异的约１２１ｂｐ大小的ＤＮＡ片段，估测每份蚊虫ＤＮＡ样本中含有３个以上子孢子或１００只蚊中含有１只阳性蚊即可得此片段，而对恶性疟原虫、食蟹猴疟原虫、约氏疟原虫及正常蚊虫ＤＮＡ不能扩增出此片段。结论：此法具有高度的敏感性和特异性。     

Concurrent salmonella bacteremia in P. vivax infection--a report of 2 cases at the Hospital for Tropical Diseases, Thailand

Malaria and concurrent bacteremia has been described in many reports, most of them with P. falciparum. Concurrent bacteremia with P. vivax infected patients is very rare. We reported 2 cases of salmonella bacteremia with P. vivax infection. Both patients presented with fever and the diagnosis of P. vivax was confirmed microscopically. The first patient presented with fever, jaundice, shock and renal failure which rarely occurs with P. vivax infection. The second patient had no clinical response after receiving standard antimalarial drugs. Hemoculture was positive for Salmonella spp in both cases. They recovered completely after appropriate antibiotics and antimalarial treatment.     

P. vivax malaria complicated by shock and ARDS

We report a case of Plasmodium vivax malaria complicated by shock and ARDS. The patient responded to oral chloroquine and primaquine and PCR was positive for P. vivax DNA and negative for P. falciparum DNA. P. vivax may cause severe complications and where the possibility of mixed infections exists, blood should be sent for PCR analysis so that mixed infections can reliably be excluded.     

Global distribution of a variant of the circumsporozoite gene of Plasmodium vivax

The global distribution of a newly described variant of the Plasmodium vivax circumsporozoite (CS) gene was determined by genetic analysis of wild isolates. Whole blood specimens were collected on filter paper from patients infected with P. vivax in South America. West Africa, and the Indian subcontinent. P. vivax DNA was released from the filter paper samples, and the CS gene was amplified by polymerase chain reaction and analyzed for genetic variation. Amplified DNA was probed with oligonucleotide probes that hybridize with the predominant CS repeat region (PV210) and the variant CS repeat region (PV247) of P. vivax. The PV247 variant was found in all three geographically diverse areas. In addition, five of six consecutive patients studied had simultaneous infection with both the predominant and variant forms of P. vivax. These findings suggest that a single-epitope vaccine based on the predominant CS domain is unlikely to be protective on even a regional basis.     

Monoclonal antibodies that distinguish Trypanosoma congolense, T. vivax and T. brucei

Monoclonal antibodies (MoAbs) were derived against in-vitro-propagated procyclic forms of Trypanosoma congolense, T. vivax, T. brucei brucei and T.b. rhodesiense in order to identify antigens for use in immunodiagnosis of African trypanosomiasis. The antibodies have been tested against procyclic and bloodstream form trypanosomes of 13 T. congolense, six T. vivax six T.b. brucei, four T.b. rhodesiense, five T.b. gambiense and three T. simiae isolates from different geographical areas by indirect immunofluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). The MoAbs raised against T.b. brucei reacted with all the brucei group of trypanosomes but not with T. congolense, T. simiae or T. vivax. Likewise, MoAbs against T. congolense reacted with T. congolense and T. simiae but not with any of the other species, while those against T. vivax reacted with T. vivax only. The antigens recognized by these MoAbs were present in lysates of bloodstream trypanosomes as well as midgut (for T. congolense and T. brucei) and epimastigote forms from infected Glossina morsitans centralis. There was no reactivity of the MoAbs with Theileria parva, Anaplasma marginale, Babesia bigemina or Plasmodium falciparum. These antibodies and the antigens they recognize should, therefore, prove useful in the development of assay systems for immunodiagnosis of African trypanosomiasis.     

Status febrilis mit Bewusstlosigkeit

Cerebral malaria with Plasmodium vivax is uncommon. Normally Plasmodium falciparum is the cause of cerebral malaria. We report about a 18 year old patient from Pakistan with a history of intermittent fever for several months. The patient recovered within a few days; however prognosis can be severe when cerebral malaria is complicating the course of Plasmodium vivax infection.     

套式PCR检测不同地理株间日疟原虫及部分SSU rRNA基因序列鉴定

为了确定扩增的特定 Ｓ Ｓ Ｕｒ Ｒ Ｎ Ａ 基因片段是否适用于不同地理株间日疟原虫的检测,在以往工作的基础上,用同一套式 Ｐ Ｃ Ｒ 体系对 ２２ 例四川和 ３０ 例云南患者及１ 例在印度感染疟疾的归国患者进行间日疟筛查,并从 Ｐ．ｖ 阳性的四川和云南血样中任取 １ 例,与 Ｐ．ｖ 阳性的印度血样分别扩增,用 Ｐ Ｃ Ｒ 产物测序,结果显示３ 者序列完全一致;与美国 Ｎ Ｃ Ｂ Ｉ的网上基因数据库比较,发现这３ 者又与萨尔瓦多等其他５ 个间日疟原虫不同地理株的小亚单位核糖体核糖核酸相应基因片段完全一致。该结果进一步证实了所扩增的片段确为目的片段,并提示该诊断体系可以适用于不同地区的间日疟原虫地理株的检测。     

Postmortem Characterization of Patients With Clinical Diagnosis of Plasmodium vivax Malaria: To What Extent Does This Parasite Kill?

Background.?Severe disease attributable to Plasmodium vivax infection is already well described worldwide; however, autopsies in these patients are scarce. Methods.?From 1996 to 2010, 19 patient deaths with a clinical diagnosis of P. vivax infection occurred in a tertiary care center in the Brazilian Amazon. Seventeen of these 19 deaths were fully autopsied. Clinical charts, macroscopic autopsy reports, and stored paraffinized tissue blocks were retrieved. Nested polymerase chain reaction was performed in paraffinized samples of spleen and lung to confirm P. vivax monoinfection. Immunohistofluorescence was used to detect P. vivax parasitized red blood cells (RBCs). Results.?Of 17 autopsies, 13 revealed that death could be attributed to P. vivax infection; in the remaining 4, acute diseases other than malaria were found to be the cause of death. The primary complication in patients in which malaria contributed to death was acute respiratory distress syndrome (ARDS) and pulmonary edema associated with the accumulation of neutrophils in the interalveolar space (6 cases). Spleen rupture (3 cases) and multiorgan dysfunction syndrome (3 cases) were the second most common complications. One child evolving with coma was also characterized, but no parasite was detected in the brain tissue. In one patient who developed ARDS and presented negative peripheral parasitemia by the time of death, scattered parasitized red blood cells were seen inside pulmonary capillaries, suggesting some sequestration in the lung. Conclusions.?In 13 of 17 deceased patients, P. vivax infection was the plausible cause of death. However, more studies are needed to understand pathogenesis related to severe disease.     

Use of PCR/DNA probes to identify circumsporozoite genotype of Plasmodium vivax in China

The paper reports the result of identifying cirumsporozoite (CS) genotype of Plasmodium vivax by using PCR/DNA probe labeled with biotin. The sensitivity of this method to detect patient blood samples was 0.2 parasite/microl and also with high specific to P. vivax. CS genes from 52 blood samples collected from patients with P. vivax in Hainan and Yunnan Provinces were amplified by PCR and 49 were positive by gel-e electrophoresis analysis, positive rate was 94%. Then the amplified CS genes further were probed with special oligoprobes (PV210 and PV247) that hybridized with the predominant CS repeat region and the variant CS repeat region. The results showed 46 (88.5%) PV210 positive and 6 (11.5%) PV247 positive; 2 hybridized with both probes. The variant genotype was present only in samples from Yunnan Province. The above results showed that the PCR/DNA probe labeled with biotin was highly sensitive and specific to P. vivax and found a CS variant genotype of P. vivax in Yunnan Province of China.     

Markers for population genetic analysis of human plasmodia species, P. falciparum and P. vivax

Present report deals with the genetic diversity existing among the field isolates of Plasmodium falciparum and P. vivax in India. Isoenzymes and molecular markers were used to analyse field isolates of P. falciparum and P. vivax. High level of length polymorphism was observed in repeat nucleotide sequences of MSP-1, MSP-2 and GLURP in P. falciparum isolates and CSP, GAM-1 and MSP-3 alpha in P. vivax isolates. In study populations a high proportion of isolates (up to 60%) were comprised of more than one genetically distinct parasite type--multiclonal. Presence of identical allelic forms of enzyme and DNA variations in different geographical areas and in different years suggest that isolates belong to a single random mating population of P. vivax and P. falciparum. Observed random combination of alleles in the field isolates suggest the unlinked nature of loci studied. Study supports the feasibility of using molecular markers for the identification of recrudescence in P. falciparum from fresh infection.     

Guillain-Barre syndrome following malaria

Two adult males were admitted with acute are flexic quadriplegia and bifacial and bulbar weakness 2 weeks after an acute episode of malaria, one due to Plasmodium falciparum infection (patient 1) and the other due to Plasmodium vivax (patient 2). Cerebrospinal fluid analysis and nerve conduction studies confirmed the diagnosis of Guillain-Barre syndrome (GBS). Patient 1 progressed to develop respiratory paralysis and required mechanical ventilation. He received intravenous immunoglobulins for the GBS and made a complete recovery in 6 weeks. A review of 11 cases of GBS (nine previously reported and the present two) revealed that eight patients had preceding falciparum malaria and three had vivax infection. All but two patients had distal symmetric sensory deficits. Paralysis was mild in seven cases (three due to P. vivax and four due to P. falciparum) and recovered completely in 2-6 weeks without any specific treatment. Four patients with falciparum malaria developed severe paralysis with respiratory failure, and three patients died. One patient who received intravenous immunoglobulins recovered completely (patient 1 in this report).